非动脉炎性前部缺血性视神经病变模型大鼠视网膜蛋白质组学定量分析

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目的:分析非动脉炎性前部缺血性视神经病变(NAION)模型大鼠视网膜蛋白质表达变化。方法:Sprague-Dawley大鼠20只,随机分为NAION动物(rNAION)模型组、单纯激光照射组、单纯光敏剂孟加拉玫瑰红(RB)组(单纯光敏剂RB组)、正常对照组,每组各为5只,均以右眼为实验眼。采用光动力疗法建立rNAION模型。建模后3 d分离各组大鼠视网膜。采用酶切法进行样本制备;非数据依赖方式采集质谱数据;SWATH定量质谱技术对数据进行定量分析,筛选差异蛋白并进行功能及相关通路分析。结果:rNAION模型组共筛选出差异蛋白184个(表达倍数大于1.5且n P<0.05 )。其中,上调蛋白99个,下调蛋白85个。胶质纤维酸性蛋白、鸟嘌呤核苷酸结合蛋白4、层粘连蛋白1、14-3-3γ蛋白YWHAG等蛋白表达增加;富含亮氨酸胶质瘤失活蛋白1、分泌载体膜蛋白5及网格蛋白外套蛋白AP180表达降低。差异蛋白主要参与神经生长、能量代谢、囊泡介导的转运、突触可塑性调节、凋亡及炎症反应等生物学进程。通路富集分析结果显示,差异蛋白主要参与磷脂酰肌醇3激酶/蛋白激酶B (PI3K/Akt)信号通路及补体和凝血酶联反应等信号通路。n 结论:NAION的发生可能通过改变能量代谢、神经生长、突触囊泡的运输及PI3K/Akt信号通路等蛋白表达,共同调控神经细胞再生及凋亡。“,”Objective:To analyze the protein expression changes in the retina of non-arteritic anterior ischemic optic neuropathy (NAION) in rats.Methods:The rat NAION (rNAION) model was established by Rose Bengal and laser. Twenty Sprague-Dawley rats were randomly divided into 4 groups, the normal control group, the laser control group, the RB injection control group, and the rNAION model group, with 5 rats in each group. The right eye was used as the experimental eye. The retina was dissected at the third day after modeling. Enzyme digestion method was used for sample preparation and data collection was performed in a non-dependent collection mode. The data were quantitatively analyzed by SWATH quantitative mass spectrometry, searching for differential proteins and performing function and pathway analysis.Results:Compared with the other three control groups, a total of 184 differential proteins were detected in the rNAION group (expression fold greater than 1.5 times and n P<0.05), including 99 up-regulated proteins and 85 down-regulated proteins. The expressions of glial fibrillary acidic protein, guanine nucleotide binding protein 4, laminin 1, 14-3-3γ protein YWHAG were increased. Whereas the expressions of Leucine-rich glioma-inactivated protein 1, secretory carrier-associated membrane protein 5, and Clathrin coat assembly protein AP180 were decreased. The differential proteins are mainly involved in biological processes such as nerve growth, energy metabolism, vesicle-mediated transport, the regulation of synaptic plasticity, apoptosis and inflammation. Pathway enrichment analysis showed that PI3K-Akt signaling pathway and complement and thrombin reaction pathway was related to the disease.n Conclusion:The protein expressions of energy metabolism, nerve growth, synaptic vesicle transport and PI3K-Akt signaling pathway can regulate the neuronal regeneration and apoptosis in NAION.
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